| 中药废水污泥群落结构解析中PCR-DGGE引物的选择与评价 |
| 摘要点击 3492 全文点击 1882 投稿时间：2010-04-12 修订日期：2010-10-01 |
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| 中文关键词 中药废水 活性污泥 16S rDNA DGGE 微生物多样性 |
| 英文关键词 traditional Chinese medicine wastewater active sludge 16S rDNA DGGE microbial diversity |
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| 中文摘要 |
| 在中药废水生物活性污泥群落DGGE解析过程中，为了探讨16S rDNA通用引物的扩增效率和对污泥细菌群落的表征能力，选用包括简并性引物在内的11对通用引物扩增16S rDNA序列的4个可变区，并应用DGGE图谱评价不同引物的扩增效率和微生物种群多样性. 结果表明，采用不同引物对进行DGGE分析时，微生物种群多样性存在着显著的差异，其中V3和V6～V8可变区分辨率较高，图谱中条带丰富，多样性较好，适合分析中药废水生物活性污泥群落的结构，而V1～V3和V3～V5可变区DGGE图谱多样性较差. 基于此，采用16S rDNA的V3和V6～V8区引物调查中药废水生物活性污泥群落都是适宜的，若考虑到序列携带更多的分类信息，建议应用V6～V8可变区引物（B968F/B1401R）；为获得更高的群落多样性信息，建议将简并和通用引物扩增的V3区PCR产物等量混合后进行DGGE分析. |
| 英文摘要 |
| In the study of traditional Chinese medicine wastewater,it was discussed for the effect of different sets of 16S rDNA universal primers on DGGE fingerprinting and microbial community diversity of aerobic and anaerobic activated sludge from one traditional Chinese medicine wastewater treatment. The genome DNA of activated sludge was isolated,and eleven sets of primers were used to amplify the four variable regions of 16S rDNA,the resolution of DGGE fingerprinting and community diversity was analyzed. The results indicated that community diversity with different sets of universal primers by DGGE was obviously different. Separated patterns of the V3 and V6-V8 regions were better than of V1-V3 and V3-V5. In the DGGE profiles,bands and diversity from V3 were most,bands and diversity from V3-V5 and V6-V8 were a little worse than those of V3. According to the length of targeted sequence and the resolution of DGGE fingerprinting,V6-V8 (B968F/B1401R) are recommended to be used to do the DGGE analysis. Mix I341F/I534R and B341F/B534R PCR product equally to make DGGE analysis can get more community diversity information. |
